The objective of this research is to determine the role of water in cancer cell division by investigating the changes in the physical state of water that occur during the synchronized HeLa cell cycle. Cells will be grown and synchronized by established cell biology methods. Samples taken during mitosis, G1, S, and G2 will be examined by nuclear magnetic resonance spectroscopy (NMR). Spin-lattice, T1, and spin-spin, T2, relaxation times of intracellular water hydrogen protons will be determined at 30 MHz and 25 degrees C by pulsed NMR techniques. Spin echo techniques will be used to determine the self diffusion coefficient of water (D) in cellular protoplasm. Percent water content and electrolytes will be determined through the cell cycle by gravimetric analysis and flame photometry. Changes in the physical state of water in the cancer cell during the cell cycle, as defined by the NMR parameters, will be correlated with the known biochemical events of the cell cycle. Methodology is proposed to study the contributions of chromatin, microtubules, and actin fibers to the cellular structure of water by macromolecular manipulation. The state of water in karyoplasts and cytoplasts of each phase of the cycle, prepared by the Cytochalasin B enucleation technique, will be examined by NMR and the relative contribution of each to the state of cellular water will be determined. These experiments will allow an evaluation of the water-macromolecular interactions characteristic of the HeLa cell cycle and will provide insights into the role that water plays in the events leading to cell division.